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  • Substrate inhibition is a common phenomenon in enzyme kinetics. We report here for the first time its study by a combination of the electrophoretically mediated microanalysis (EMMA) methodology with a partial filling technique. In this set-up, the part of capillary is filled with the buffer best for the enzymatic reaction whereas, the rest of the capillary is filled with the background electrolyte optimal for separation of substrates and products. In the case of haloalkane dehalogenase, a model enzyme selected for this study, the enzymatic reaction was performed in 20 mM glycine buffer (pH 8.6) whereas 20 mM beta-alanine - hydrochloric acid buffer (pH 3.5) was used as a background electrolyte in combination with direct detection at 200 nm. The whole study was performed on poorly soluble brominated substrate - 1,2-dibromoethane.
  • Substrate inhibition is a common phenomenon in enzyme kinetics. We report here for the first time its study by a combination of the electrophoretically mediated microanalysis (EMMA) methodology with a partial filling technique. In this set-up, the part of capillary is filled with the buffer best for the enzymatic reaction whereas, the rest of the capillary is filled with the background electrolyte optimal for separation of substrates and products. In the case of haloalkane dehalogenase, a model enzyme selected for this study, the enzymatic reaction was performed in 20 mM glycine buffer (pH 8.6) whereas 20 mM beta-alanine - hydrochloric acid buffer (pH 3.5) was used as a background electrolyte in combination with direct detection at 200 nm. The whole study was performed on poorly soluble brominated substrate - 1,2-dibromoethane. (en)
Title
  • Study of substrate inhibition by electrophoretically mediated microanalysis in partially filled capillary.
  • Study of substrate inhibition by electrophoretically mediated microanalysis in partially filled capillary. (en)
skos:prefLabel
  • Study of substrate inhibition by electrophoretically mediated microanalysis in partially filled capillary.
  • Study of substrate inhibition by electrophoretically mediated microanalysis in partially filled capillary. (en)
skos:notation
  • RIV/00216224:14310/07:00020038!RIV10-MSM-14310___
http://linked.open...avai/riv/aktivita
http://linked.open...avai/riv/aktivity
  • P(GA203/06/0047), P(LC06023), Z(MSM0021622413)
http://linked.open...iv/cisloPeriodika
  • 1-2
http://linked.open...vai/riv/dodaniDat
http://linked.open...aciTvurceVysledku
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  • 453316
http://linked.open...ai/riv/idVysledku
  • RIV/00216224:14310/07:00020038
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  • CE; EMMA substrate; inhibition; haloalkane dehalogenase (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • NL - Nizozemsko
http://linked.open...ontrolniKodProRIV
  • [B515974CE304]
http://linked.open...i/riv/nazevZdroje
  • Journal of Chromatography A
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http://linked.open...v/svazekPeriodika
  • 1150
http://linked.open...iv/tvurceVysledku
  • Chaloupková, Radka
  • Němec, Tomáš
  • Glatz, Zdeněk
  • Papežová, Kateřina
http://linked.open...n/vavai/riv/zamer
issn
  • 0021-9673
number of pages
http://localhost/t...ganizacniJednotka
  • 14310
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